通过时间分辨荧光免疫测定法检测低分子量半胱氨酸蛋白酶抑制剂

Detection of low molecular weight cysteine proteinase inhibitors by time-resolved fluoroimmunoassay.

作者信息

Joronen I, Hopsu-Havu V K, Manninen M, Rinne A, Järvinen M, Halonen P

出版信息

J Immunol Methods. 1986 Feb 12;86(2):243-7. doi: 10.1016/0022-1759(86)90460-6.

Abstract

A time-resolved fluoroimmunoassay was developed for the detection of 3 human low molecular weight cysteine proteinase inhibitors, ACPI (cystatin A), NCPI (cystatin B), and gamma-trace (cystatin C). Polystyrene tubes or polystyrene microtitration strips were used as solid phase. The rabbit anti-inhibitor immunoglobulins were used as the capture antibody, and, when labelled with europium, also as the detector antibody. The threshold sensitivity of the tests was 0.1 ng/ml for NCPI and 1 ng/ml for the others. All the 3 cysteine proteinase inhibitors, ACPI, NCPI, and gamma-trace, were detected in pooled serum samples of patients with kidney failure. gamma-Trace seemed to be quantitatively the major and ACPI the minor inhibitor. No other low molecular mass cysteine proteinase inhibitor was detected after isoelectric focusing of the 12 kDa area of gel filtered human serum.

摘要

开发了一种时间分辨荧光免疫分析法，用于检测3种人低分子量半胱氨酸蛋白酶抑制剂，即ACPI（胱抑素A）、NCPI（胱抑素B）和γ-微球蛋白（胱抑素C）。使用聚苯乙烯管或聚苯乙烯微量滴定条作为固相。兔抗抑制剂免疫球蛋白用作捕获抗体，用铕标记后也用作检测抗体。检测的阈值灵敏度对于NCPI为0.1 ng/ml，对于其他两种为1 ng/ml。在肾衰竭患者的混合血清样本中检测到了所有3种半胱氨酸蛋白酶抑制剂，即ACPI、NCPI和γ-微球蛋白。γ-微球蛋白似乎在数量上是主要的抑制剂，而ACPI是次要的抑制剂。在对凝胶过滤的人血清12 kDa区域进行等电聚焦后，未检测到其他低分子量半胱氨酸蛋白酶抑制剂。